Spinach hybrid svvc8155 and parents thereof

ABSTRACT

The invention provides seed and plants of spinach hybrid SVVC8155 and the parent lines thereof. The invention thus relates to the plants, seeds and tissue cultures of spinach hybrid SVVC8155 and the parent lines thereof, and to methods for producing a spinach plant produced by crossing such plants with themselves or with another spinach plant, such as a plant of another genotype. The invention further relates to seeds and plants produced by such crossing. The invention further relates to parts of such plants, including the leaf and gametes of such plants.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the priority of U.S. Provisional Appl. Ser. No.62/465,676, filed Mar. 1, 2017, the entire disclosure of which isincorporated herein by reference.

FIELD OF THE INVENTION

The field of the invention relates to plant breeding and, morespecifically, to the development of spinach hybrid SVVC8155 and parentspinach line SSBS012-1181F.

BACKGROUND OF THE INVENTION

The goal of vegetable breeding is to combine various desirable traits ina single variety/hybrid. Such desirable traits may include any traitdeemed beneficial by a grower and/or consumer, including greater yield,resistance to insects or pests, tolerance to environmental stress,better agronomic quality, higher nutritional value, growth rate andfruit properties.

Breeding techniques take advantage of a plant's method of pollination.There are two general methods of pollination: a plant self-pollinates ifpollen from one flower is transferred to the same or another flower ofthe same plant or plant variety. A plant cross-pollinates if pollencomes to it from a flower of a different plant variety.

Plants that have been self-pollinated and selected for type over manygenerations become homozygous at almost all gene loci and produce auniform population of true breeding progeny, a homozygous plant. A crossbetween two such homozygous plants of different genotypes produces auniform population of hybrid plants that are heterozygous for many geneloci. Conversely, a cross of two plants each heterozygous at a number ofloci produces a population of hybrid plants that differ genetically andare not uniform. The resulting non-uniformity makes performanceunpredictable.

The development of uniform varieties requires the development ofhomozygous inbred plants, the crossing of these inbred plants, and theevaluation of the crosses. Pedigree breeding and recurrent selection areexamples of breeding methods that have been used to develop inbredplants from breeding populations. Those breeding methods combine thegenetic backgrounds from two or more plants or various other broad-basedsources into breeding pools from which new lines and hybrids derivedtherefrom are developed by selfing and selection of desired phenotypes.The new lines and hybrids are evaluated to determine which of those havecommercial potential.

SUMMARY OF THE INVENTION

In one aspect, the present invention provides a spinach plant of thehybrid designated SVVC8155, the spinach line SSBS012-1181F, or thespinach line SSB-S015-1249M. Also provided are spinach plants having allthe physiological and morphological characteristics of such a plant.Parts of these spinach plants are also provided, for example, includingpollen, an ovule, scion, a rootstock, a fruit, and a cell of the plant.

In another aspect of the invention, a plant of spinach hybrid SVVC8155and/or spinach line SSBS012-1181F comprising an added heritable trait isprovided. The heritable trait may comprise a genetic locus that is, forexample, a dominant or recessive allele. In one embodiment of theinvention, a plant of spinach hybrid SVVC8155 and/or spinach lineSSBS012-1181F is defined as comprising a single locus conversion. Inspecific embodiments of the invention, an added genetic locus confersone or more traits such as, for example, herbicide tolerance, insectresistance, disease resistance, and modified carbohydrate metabolism. Infurther embodiments, the trait may be conferred by a naturally occurringgene introduced into the genome of a line by backcrossing, a natural orinduced mutation, or a transgene introduced through genetictransformation techniques into the plant or a progenitor of any previousgeneration thereof. When introduced through transformation, a geneticlocus may comprise one or more genes integrated at a single chromosomallocation.

The invention also concerns the seed of spinach hybrid SVVC8155 and/orspinach line SSBS012-1181F. The spinach seed of the invention may beprovided as an essentially homogeneous population of spinach seed ofspinach hybrid SVVC8155 and/or spinach line SSBS012-1181F. Essentiallyhomogeneous populations of seed are generally free from substantialnumbers of other seed. Therefore, seed of hybrid SVVC8155 and/or spinachline SSBS012-1181F may be defined as forming at least about 97% of thetotal seed, including at least about 98%, 99%, or more of the seed. Theseed population may be separately grown to provide an essentiallyhomogeneous population of spinach plants designated SVVC8155 and/orspinach line SSBS012-1181F.

In yet another aspect of the invention, a tissue culture of regenerablecells of a spinach plant of hybrid SVVC8155 and/or spinach lineSSBS012-1181F is provided. The tissue culture will preferably be capableof regenerating spinach plants capable of expressing all of thephysiological and morphological characteristics of the starting plantand of regenerating plants having substantially the same genotype as thestarting plant. Examples of some of the physiological and morphologicalcharacteristics of the hybrid SVVC8155 and/or spinach line SSBS012-1181Finclude those traits set forth in the tables herein. The regenerablecells in such tissue cultures may be derived, for example, from embryos,meristems, cotyledons, pollen, leaves, anthers, roots, root tips,pistils, flowers, seed, and stalks. Still further, the present inventionprovides spinach plants regenerated from a tissue culture of theinvention, the plants having all the physiological and morphologicalcharacteristics of hybrid SVVC8155 and/or spinach line SSBS012-1181F.

In still yet another aspect of the invention, processes are provided forproducing spinach seeds, plants, and fruit, which processes generallycomprise crossing a first parent spinach plant with a second parentspinach plant, wherein at least one of the first or second parentspinach plants is a plant of spinach lines SSBS012-1181F orSSB-S015-1249M. These processes may be further exemplified as processesfor preparing hybrid spinach seed or plants, wherein a first spinachplant is crossed with a second spinach plant of a different, distinctgenotype to provide a hybrid that has, as one of its parents, a plant ofspinach line SSBS012-1181F or SSB-S015-1249M. In these processes,crossing will result in the production of seed. The seed productionoccurs regardless of whether the seed is collected or not.

In one embodiment of the invention, the first step in “crossing”comprises planting seeds of a first and second parent spinach plant,often in proximity so that pollination will occur for example, mediatedby insect vectors. Alternatively, pollen can be transferred manually.Where the plant is self-pollinated, pollination may occur without theneed for direct human intervention other than plant cultivation.

A second step may comprise cultivating or growing the seeds of first andsecond parent spinach plants into plants that bear flowers. A third stepmay comprise preventing self-pollination of the plants, such as byemasculating the flowers (i.e., killing or removing the pollen).

A fourth step for a hybrid cross may comprise cross-pollination betweenthe first and second parent spinach plants. Yet another step comprisesharvesting the seeds from at least one of the parent spinach plants. Theharvested seed can be grown to produce a spinach plant or hybrid spinachplant.

The present invention also provides the spinach seeds and plantsproduced by a process that comprises crossing a first parent spinachplant with a second parent spinach plant, wherein at least one of thefirst or second parent spinach plants is a plant of spinach hybridSVVC8155 and/or spinach line SSBS012-1181F. In one embodiment of theinvention, spinach seed and plants produced by the process are firstgeneration (F₁) hybrid spinach seed and plants produced by crossing aplant in accordance with the invention with another, distinct plant. Thepresent invention further contemplates plant parts of such an F₁ hybridspinach plant and methods of use thereof. Therefore, certain exemplaryembodiments of the invention provide an F₁ hybrid spinach plant and seedthereof.

In still yet another aspect, the present invention provides a method ofproducing a plant derived from hybrid SVVC8155 and/or spinach lineSSBS012-1181F, the method comprising the steps of: (a) preparing aprogeny plant derived from hybrid SVVC8155 and/or spinach lineSSBS012-1181F, wherein said preparing comprises crossing a plant of thehybrid SVVC8155 and/or spinach line SSBS012-1181F with a second plant;and (b) crossing the progeny plant with itself or a second plant toproduce a seed of a progeny plant of a subsequent generation. In furtherembodiments, the method may additionally comprise: (c) growing a progenyplant of a subsequent generation from said seed of a progeny plant of asubsequent generation and crossing the progeny plant of a subsequentgeneration with itself or a second plant; and repeating the steps for anadditional 3-10 generations to produce a plant derived from hybridSVVC8155 and/or spinach line SSBS012-1181F. The plant derived fromhybrid SVVC8155 and/or spinach line SSBS012-1181F may be an inbred line,and the aforementioned repeated crossing steps may be defined ascomprising sufficient inbreeding to produce the inbred line. In themethod, it may be desirable to select particular plants resulting fromstep (c) for continued crossing according to steps (b) and (c). Byselecting plants having one or more desirable traits, a plant derivedfrom hybrid SVVC8155 and/or spinach line SSBS012-1181F is obtained whichpossesses some of the desirable traits of the line/hybrid as well aspotentially other selected traits.

In certain embodiments, the present invention provides a method ofproducing food or feed comprising: (a) obtaining a plant of spinachhybrid SVVC8155 and/or spinach line SSBS012-1181F, wherein the plant hasbeen cultivated to maturity and (b) collecting at least one spinach fromthe plant.

In still yet another aspect of the invention, the genetic complement ofspinach hybrid SVVC8155 and/or spinach line SSBS012-1181F is provided.The phrase “genetic complement” is used to refer to the aggregate ofnucleotide sequences, the expression of which sequences defines thephenotype of, in the present case, a spinach plant, or a cell or tissueof that plant. A genetic complement thus represents the genetic make-upof a cell, tissue or plant, and a hybrid genetic complement representsthe genetic make-up of a hybrid cell, tissue or plant. The inventionthus provides spinach plant cells that have a genetic complement inaccordance with the spinach plant cells disclosed herein, and seeds andplants containing such cells.

Plant genetic complements may be assessed by genetic marker profiles,and by the expression of phenotypic traits that are characteristic ofthe expression of the genetic complement, e.g., isozyme typing profiles.It is understood that hybrid SVVC8155 and/or spinach line SSBS012-1181Fcould be identified by any of the many well-known techniques such as,for example, Simple Sequence Length Polymorphisms (SSLPs) (Williams etal., Nucleic Acids Res., 1 8:6531-6535, 1990), Randomly AmplifiedPolymorphic DNAs (RAPDs), DNA Amplification Fingerprinting (DAF),Sequence Characterized Amplified Regions (SCARs), Arbitrary PrimedPolymerase Chain Reaction (AP-PCR), Amplified Fragment LengthPolymorphisms (AFLPs) (EP 534 858, specifically incorporated herein byreference in its entirety), and Single Nucleotide Polymorphisms (SNPs)(Wang et al., Science 280:1077-1082, 1998).

In still yet another aspect, the present invention provides hybridgenetic complements, as represented by spinach plant cells, tissues,plants, and seeds, formed by the combination of a haploid geneticcomplement of a spinach plant of the invention with a haploid geneticcomplement of a second spinach plant, preferably, another, distinctspinach plant. In another aspect, the present invention provides aspinach plant regenerated from a tissue culture that comprises a hybridgenetic complement of this invention.

Any embodiment discussed herein with respect to one aspect of theinvention applies to other aspects of the invention as well, unlessspecifically noted.

The term “about” is used to indicate that a value includes the standarddeviation of the mean for the device or method being employed todetermine the value. The use of the term “or” in the claims is used tomean “and/or” unless explicitly indicated to refer to alternatives onlyor the alternatives are mutually exclusive. When used in conjunctionwith the word “comprising” or other open language in the claims, thewords “a” and “an” denote “one or more,” unless specifically notedotherwise. The terms “comprise,” “have” and “include” are open-endedlinking verbs. Any forms or tenses of one or more of these verbs, suchas “comprises,” “comprising,” “has,” “having,” “includes” and“including,” are also open-ended. For example, any method that“comprises,” “has” or “includes” one or more steps is not limited topossessing only those one or more steps and also covers other unlistedsteps. Similarly, any plant that “comprises,” “has” or “includes” one ormore traits is not limited to possessing only those one or more traitsand covers other unlisted traits.

Other objects, features, and advantages of the present invention willbecome apparent from the following detailed description. It should beunderstood, however, that the detailed description and any specificexamples provided, while indicating specific embodiments of theinvention, are given by way of illustration only, since various changesand modifications within the spirit and scope of the invention willbecome apparent to those skilled in the art from this detaileddescription.

DETAILED DESCRIPTION OF THE INVENTION

The invention provides methods and compositions relating to plants,seeds, and derivatives of spinach hybrid SVVC8155, spinach lineSSBS012-1181F, or spinach line SSB-S015-1249M.

Spinach hybrid SVVC8155, also known as 15-S0-SMT-5587, is a slowmaturing variety developed for the late spring and early autumn growingseasons. The hybrid has medium, dark, semi-oval/pointed, erect leavesand resistance to downy mildew (Peronospora farinose f. sp. spinaciaeraces 1-15).

Parent line SSBS012-1181F is a mid-late bolting, female-floweringvariety with very dark leaves, a compact plant habit, and resistance todowny mildew (Peronospora farinose f. sp. spinaciae races 1-10, 15).

Parent line SSB-S015-1249M is a mid-late bolting, male-flowering varietywith medium-dark, oval, semi-savoy leaves; some reverse cupping; andresistance to downy mildew (Peronospora farinose f. sp. spinaciae races1-6, 8, 9, 11-15).

A. ORIGIN AND BREEDING HISTORY OF SPINACH HYBRID SVVC8155

Hybrid SVVC8155 was developed by crossing line SSBS012-1181F with lineSSB-S015-1249M. The parent lines are uniform and stable, as is a hybridproduced therefrom. A small percentage of variants can occur withincommercially acceptable limits for almost any characteristic during thecourse of repeated multiplication. However no variants are expected.

B. PHYSIOLOGICAL AND MORPHOLOGICAL CHARACTERISTICS OF SPINACH HYBRIDSVVC8155 AND SPINACH LINE SSBS012-1181F

In accordance with one aspect of the present invention, there isprovided a plant having the physiological and morphologicalcharacteristics of spinach hybrid SVVC8155 and the parent lines thereof.A description of the physiological and morphological characteristics ofsuch plants is presented in Tables 1-2.

TABLE 1 Physiological and Morphological Characteristics of HybridSVVC8155 CHARACTERISTIC SVVC8155 Boeing Ploidy diploid diploid SeedlingCotyledon Length (mm) 64.8 67.9 Length medium medium Width (mm)  6.6 6.4 Tip pointed pointed Color medium green medium green RHS color chartvalue 146b 144a Leaf (First Foliage Leaves) Shape (first foliage leaves)ovate ovate Base (first foliage leaves) V-shape V-shape Tip (firstfoliage leaves) round-pointed round-pointed Margin (first foliageleaves) flat flat Upper surface color medium green medium green [GiantNobel] [Giant Nobel] RHS color chart value 146a 146a Lower surface color(compared lighter lighter with upper surface) (first foliage leaves) RHScolor chart value 146b 146b Maturity Growth rate slow slow [Norgreen][Norgreen] Days from planting to prime 53   53   market stage Plant(Prime Market Stage) Habit flat flat [Vivoflay] [Vivoflay] Size mediummedium Spread (cm) 39.7 40.9 Height (cm) 12.1 13.2 Leaf (Prime MarketStage) Surface smooth smooth [Vivoflay] [Viroflay] Anthocyanincoloration of petioles absent absent and veins Blade intensity of greencolor medium medium Blade blistering medium medium Blade lobing mediumabsent or very weak-weak Petiole attitude horizontal horizontal Petiolelength medium medium Blade attitude semi-erect horizontal Leaf shapeovate ovate Blade shape (excluding basal broad ovate medium ovate lobes)Blade curving of margin flat incurved Blade shape of apex roundedrounded Blade shape in longitudinal section concave flat Base lobedlobed Tip round round Margin flat slightly curved Upper surface colormedium green medium green RHS color chart value  146a+ 146a Lowersurface color (compared lighter lighter with upper surface) RHS colorchart value 146b 146b Luster glossy glossy Blade size large largePetiole color medium green medium green RHS color chart value 144a 144aPetiole red pigmentation absent absent petiole length to the blade (cm)8.7 cm 10.9 cm Petiole diameter (mm)  8.0 mm   7.0 mm Petiole diametermedium medium Seed Stalk Development Start of bolting (10% of plants)late late Time of start of bolting (for spring late-very late late-verylate sown crop, 15% of plants) Height of stalk (cm) 105.0 cm  94.0 cmFlowering plants: Plants that are 0-10%  0-10% female Proportion offemale plants absent or very — low Flowering plants: Plants that are0-10% 91-100% male Proportion of male plants absent or very absent orvery low low Flowering plants: Plants that are 91-100%  91-100%monoecious Proportion of monoecious plants very high very high SeedSurface smooth smooth Spines (harvested seed) absent absent[Resistoflay] [Resistoflay] *These are typical values. Values may varydue to environment. Other values that are substantially equivalent arealso within the scope of the invention.

TABLE 2 Physiological and Morphological Characteristics of LineSSBS012-1181F CHARACTERISTIC SSBS012-1181F OMB-66-1108D Ploidy diploiddiploid Seedling Cotyledon Length (mm) 72.1 76.1 Length medium mediumWidth (mm)  7.0  7.6 Tip pointed pointed Color dark green dark green RHScolor chart value 146a 146a Leaf (First Foliage Leaves) Shape (firstfoliage leaves) elliptic elliptic Base (first foliage leaves) V-shapeV-shape Tip (first foliage leaves) round-pointed round-pointed Margin(first foliage leaves) flat flat Upper surface color medium green darkgreen [Giant Nobel] [Long Standing Bloomsdale] RHS color chart value146a 147a Lower Surface Color (compared lighter lighter with uppersurface) (first foliage leaves) RHS color chart value 146b 147b MaturityGrowth rate slow slow [Norgreen] [Norgreen] Days from planting to prime54   48   market stage Plant (Prime Market Stage) Habit semi-erectsemi-erect [Long Standing [Long Standing Bloomsdale] Bloomsdale] Sizemedium medium Spread (cm) 44.0 42.2 Height (cm) 12.0 11.8 Leaf (PrimeMarket Stage) Surface smooth smooth [Vivoflay] [Viroflay] Anthocyanincoloration of petioles absent absent and veins Blade intensity of greencolor dark dark-very dark Blade blistering weak weak Blade lobing weakweak Petiole attitude semi-erect/ semi-erect horizontal Petiole lengthmedium medium Blade attitude horizontal semi-erect Leaf shape ovateovate Blade shape (excluding basal medium ovate medium ovate lobes)Blade curving of margin flat flat Blade shape of apex obtuse obtuseBlade shape in longitudinal section flat flat Base lobed lobed Tipround-pointed round-pointed Margin flat flat Upper surface color darkgreen dark green RHS color chart value 147a  147a+ Lower surface color(compared lighter lighter with upper surface) RHS color chart value 147b147b Luster glossy glossy Blade size large medium Petiole color mediumgreen medium green RHS color chart value 144a 144a Petiole redpigmentation absent absent petiole length to the blade (cm)  9.1  9.0Petiole diameter (mm)  9.5  9.0 Petiole diameter medium medium SeedStalk Development Start of bolting (10% of plants) late late Time ofstart of bolting (for spring late-very late late sown crop, 15% ofplants) Height of stalk (cm) 76.6 84.0 Flowering plants: Plants that are0-10% 36-65% female Proportion of female plants absent or very mediumlow Flowering plants: Plants that are 0-10% 36-65% male Proportion ofmale plants absent or very medium low Flowering plants: Plants that are91-100%   1-10% monoecious Proportion of monoecious plants very highabsent or very low Seed Surface smooth smooth Spines (harvested seed)absent absent [Resistoflay] [Resistoflay] *These are typical values.Values may vary due to environment. Other values that are substantiallyequivalent are also within the scope of the invention.

C. BREEDING SPINACH PLANTS

One aspect of the current invention concerns methods for producing seedof spinach hybrid SVVC8155 involving crossing spinach linesSSBS012-1181F and SSB-S015-1249M. Alternatively, in other embodiments ofthe invention, hybrid SVVC8155, line SSBS012-1181F, or lineSSB-S015-1249M may be crossed with itself or with any second plant. Suchmethods can be used for propagation of hybrid SVVC8155 and/or thespinach lines SSBS012-1181F and SSB-S015-1249M, or can be used toproduce plants that are derived from hybrid SVVC8155 and/or the spinachlines SSBS012-1181F and SSB-S015-1249M. Plants derived from hybridSVVC8155 and/or the spinach lines SSBS012-1181F and SSB-S015-1249M maybe used, in certain embodiments, for the development of new spinachvarieties.

The development of new varieties using one or more starting varieties iswell-known in the art. In accordance with the invention, novel varietiesmay be created by crossing hybrid SVVC8155 followed by multiplegenerations of breeding according to such well-known methods. Newvarieties may be created by crossing with any second plant. In selectingsuch a second plant to cross for the purpose of developing novel lines,it may be desired to choose those plants which either themselves exhibitone or more selected desirable characteristics or which exhibit thedesired characteristic(s) when in hybrid combination. Once initialcrosses have been made, inbreeding and selection take place to producenew varieties. For development of a uniform line, often five or moregenerations of selfing and selection are involved.

Uniform lines of new varieties may also be developed by way ofdouble-haploids. This technique allows the creation of true breedinglines without the need for multiple generations of selfing andselection. In this manner true breeding lines can be produced in aslittle as one generation. Haploid embryos may be produced frommicrospores, pollen, anther cultures, or ovary cultures. The haploidembryos may then be doubled autonomously, or by chemical treatments(e.g., colchicine treatment). Alternatively, haploid embryos may begrown into haploid plants and treated to induce chromosome doubling. Ineither case, fertile homozygous plants are obtained. In accordance withthe invention, any of such techniques may be used in connection with aplant of the invention and progeny thereof to achieve a homozygous line.

Backcrossing can also be used to improve an inbred plant. Backcrossingtransfers a specific desirable trait from one inbred or non-inbredsource to an inbred that lacks that trait. This can be accomplished, forexample, by first crossing a superior inbred (A) (recurrent parent) to adonor inbred (non-recurrent parent), which carries the appropriate locusor loci for the trait in question. The progeny of this cross are thenmated back to the superior recurrent parent (A) followed by selection inthe resultant progeny for the desired trait to be transferred from thenon-recurrent parent. After five or more backcross generations withselection for the desired trait, the progeny have the characteristicbeing transferred, but are like the superior parent for most or almostall other loci. The last backcross generation would be selfed to givepure breeding progeny for the trait being transferred.

The plants of the present invention are particularly well suited for thedevelopment of new lines based on the elite nature of the geneticbackground of the plants. In selecting a second plant to cross withSVVC8155 and/or spinach lines SSBS012-1181F and SSB-S015-1249M for thepurpose of developing novel spinach lines, it will typically bepreferred to choose those plants which either themselves exhibit one ormore selected desirable characteristics or which exhibit the desiredcharacteristic(s) when in hybrid combination. Examples of desirabletraits may include, in specific embodiments, high seed yield, high seedgermination, seedling vigor, high fruit yield, disease tolerance orresistance, and adaptability for soil and climate conditions.Consumer-driven traits, such as a fruit shape, color, texture, and tasteare other examples of traits that may be incorporated into new lines ofspinach plants developed by this invention.

D. FURTHER EMBODIMENTS OF THE INVENTION

In certain aspects of the invention, plants described herein areprovided modified to include at least a first desired heritable trait.Such plants may, in one embodiment, be developed by a plant breedingtechnique called backcrossing, wherein essentially all of themorphological and physiological characteristics of a variety arerecovered in addition to a genetic locus transferred into the plant viathe backcrossing technique. The term single locus converted plant asused herein refers to those spinach plants which are developed by aplant breeding technique called backcrossing, wherein essentially all ofthe morphological and physiological characteristics of a variety arerecovered in addition to the single locus transferred into the varietyvia the backcrossing technique. By essentially all of the morphologicaland physiological characteristics, it is meant that the characteristicsof a plant are recovered that are otherwise present when compared in thesame environment, other than an occasional variant trait that mightarise during backcrossing or direct introduction of a transgene.

Backcrossing methods can be used with the present invention to improveor introduce a characteristic into the present variety. The parentalspinach plant which contributes the locus for the desired characteristicis termed the nonrecurrent or donor parent. This terminology refers tothe fact that the nonrecurrent parent is used one time in the backcrossprotocol and therefore does not recur. The parental spinach plant towhich the locus or loci from the nonrecurrent parent are transferred isknown as the recurrent parent as it is used for several rounds in thebackcrossing protocol.

In a typical backcross protocol, the original variety of interest(recurrent parent) is crossed to a second variety (nonrecurrent parent)that carries the single locus of interest to be transferred. Theresulting progeny from this cross are then crossed again to therecurrent parent and the process is repeated until a spinach plant isobtained wherein essentially all of the morphological and physiologicalcharacteristics of the recurrent parent are recovered in the convertedplant, in addition to the single transferred locus from the nonrecurrentparent.

The selection of a suitable recurrent parent is an important step for asuccessful backcrossing procedure. The goal of a backcross protocol isto alter or substitute a single trait or characteristic in the originalvariety. To accomplish this, a single locus of the recurrent variety ismodified or substituted with the desired locus from the nonrecurrentparent, while retaining essentially all of the rest of the desiredgenetic, and therefore the desired physiological and morphologicalconstitution of the original variety. The choice of the particularnonrecurrent parent will depend on the purpose of the backcross; one ofthe major purposes is to add some commercially desirable trait to theplant. The exact backcrossing protocol will depend on the characteristicor trait being altered and the genetic distance between the recurrentand nonrecurrent parents. Although backcrossing methods are simplifiedwhen the characteristic being transferred is a dominant allele, arecessive allele, or an additive allele (between recessive anddominant), may also be transferred. In this instance it may be necessaryto introduce a test of the progeny to determine if the desiredcharacteristic has been successfully transferred.

In one embodiment, progeny spinach plants of a backcross in which aplant described herein is the recurrent parent comprise (i) the desiredtrait from the non-recurrent parent and (ii) all of the physiologicaland morphological characteristics of spinach the recurrent parent asdetermined at the 5% significance level when grown in the sameenvironmental conditions.

New varieties can also be developed from more than two parents. Thetechnique, known as modified backcrossing, uses different recurrentparents during the backcrossing. Modified backcrossing may be used toreplace the original recurrent parent with a variety having certain moredesirable characteristics or multiple parents may be used to obtaindifferent desirable characteristics from each.

With the development of molecular markers associated with particulartraits, it is possible to add additional traits into an established germline, such as represented here, with the end result being substantiallythe same base germplasm with the addition of a new trait or traits.Molecular breeding, as described in Moose and Mumm, 2008 (Plant Physiol.147:969-977), for example, and elsewhere, provides a mechanism forintegrating single or multiple traits or QTL into an elite line. Thismolecular breeding-facilitated movement of a trait or traits into anelite line may encompass incorporation of a particular genomic fragmentassociated with a particular trait of interest into the elite line bythe mechanism of identification of the integrated genomic fragment withthe use of flanking or associated marker assays. In the embodimentrepresented here, one, two, three, or four genomic loci, for example,may be integrated into an elite line via this methodology. When thiselite line containing the additional loci is further crossed withanother parental elite line to produce hybrid offspring, it is possibleto then incorporate at least eight separate additional loci into thehybrid. These additional loci may confer, for example, such traits as adisease resistance or a fruit quality trait. In one embodiment, eachlocus may confer a separate trait. In another embodiment, loci may needto be homozygous and exist in each parent line to confer a trait in thehybrid. In yet another embodiment, multiple loci may be combined toconfer a single robust phenotype of a desired trait.

Many single locus traits have been identified that are not regularlyselected for in the development of a new inbred but that can be improvedby backcrossing techniques. Single locus traits may or may not betransgenic; examples of these traits include, but are not limited to,herbicide resistance, resistance to bacterial, fungal, or viral disease,insect resistance, modified fatty acid or carbohydrate metabolism, andaltered nutritional quality. These comprise genes generally inheritedthrough the nucleus.

Direct selection may be applied where the single locus acts as adominant trait. For this selection process, the progeny of the initialcross are assayed for viral resistance and/or the presence of thecorresponding gene prior to the backcrossing. Selection eliminates anyplants that do not have the desired gene and resistance trait, and onlythose plants that have the trait are used in the subsequent backcross.This process is then repeated for all additional backcross generations.

Selection of spinach plants for breeding is not necessarily dependent onthe phenotype of a plant and instead can be based on geneticinvestigations. For example, one can utilize a suitable genetic markerwhich is closely genetically linked to a trait of interest. One of thesemarkers can be used to identify the presence or absence of a trait inthe offspring of a particular cross, and can be used in selection ofprogeny for continued breeding. This technique is commonly referred toas marker assisted selection. Any other type of genetic marker or otherassay which is able to identify the relative presence or absence of atrait of interest in a plant can also be useful for breeding purposes.Procedures for marker assisted selection are well-known in the art. Suchmethods will be of particular utility in the case of recessive traitsand variable phenotypes, or where conventional assays may be moreexpensive, time consuming or otherwise disadvantageous. Types of geneticmarkers which could be used in accordance with the invention include,but are not necessarily limited to, Simple Sequence Length Polymorphisms(SSLPs) (Williams et al., Nucleic Acids Res. 18:6531-6535, 1990),Randomly Amplified Polymorphic DNAs (RAPDs), DNA AmplificationFingerprinting (DAF), Sequence Characterized Amplified Regions (SCARs),Arbitrary Primed Polymerase Chain Reaction (AP-PCR), Amplified FragmentLength Polymorphisms (AFLPs) (EP 534 858, specifically incorporatedherein by reference in its entirety), and Single NucleotidePolymorphisms (SNPs) (Wang et al., Science 280:1077-1082, 1998).

E. PLANTS DERIVED BY GENETIC ENGINEERING

Many useful traits that can be introduced by backcrossing, as well asdirectly into a plant, are those which are introduced by moleculargenetic methods. Such methods include, but are not limited to, variousplant transformation techniques and methods for site-specificrecombination, the use of which are well-known in the art and include,for example, the CRISPR-Cas system, zinc-finger nucleases (ZFNs), andtranscription activator-like effector nucleases (TALENs), among others.

In one embodiment of the invention, genetic transformation may be usedto insert a selected transgene into a plant of the invention or may,alternatively, be used for the preparation of transgenes which can beintroduced by backcrossing. Methods for the transformation of plantsthat are well-known to those of skill in the art and applicable to manycrop species include, but are not limited to, electroporation,microprojectile bombardment, Agrobacterium-mediated transformation, anddirect DNA uptake by protoplasts.

To effect transformation by electroporation, one may employ eitherfriable tissues, such as a suspension culture of cells or embryogeniccallus or alternatively one may transform immature embryos or otherorganized tissue directly. In this technique, one would partiallydegrade the cell walls of the chosen cells by exposing them topectin-degrading enzymes (pectolyases) or mechanically wound tissues ina controlled manner.

An efficient method for delivering transforming DNA segments to plantcells is microprojectile bombardment. In this method, particles arecoated with nucleic acids and delivered into cells by a propellingforce. Exemplary particles include those comprised of tungsten,platinum, and preferably, gold. For the bombardment, cells in suspensionare concentrated on filters or solid culture medium. Alternatively,immature embryos or other target cells may be arranged on solid culturemedium. The cells to be bombarded are positioned at an appropriatedistance below the macroprojectile stopping plate.

An illustrative embodiment of a method for delivering DNA into plantcells by acceleration is the Biolistics Particle Delivery System, whichcan be used to propel particles coated with DNA or cells through ascreen, such as a stainless steel or Nytex screen, onto a surfacecovered with target cells. The screen disperses the particles so thatthey are not delivered to the recipient cells in large aggregates.Microprojectile bombardment techniques are widely applicable, and may beused to transform virtually any plant species.

Agrobacterium-mediated transfer is another widely applicable system forintroducing gene loci into plant cells. An advantage of the technique isthat DNA can be introduced into whole plant tissues, thereby bypassingthe need for regeneration of an intact plant from a protoplast. ModernAgrobacterium transformation vectors are capable of replication in E.coli as well as Agrobacterium, allowing for convenient manipulations(Klee et al., Nat. Biotechnol. 3(7):637-642, 1985). Moreover, recenttechnological advances in vectors for Agrobacterium-mediated genetransfer have improved the arrangement of genes and restriction sites inthe vectors to facilitate the construction of vectors capable ofexpressing various polypeptide coding genes. The vectors described haveconvenient multi-linker regions flanked by a promoter and apolyadenylation site for direct expression of inserted polypeptidecoding genes. Additionally, Agrobacterium containing both armed anddisarmed Ti genes can be used for transformation.

In those plant strains where Agrobacterium-mediated transformation isefficient, it is the method of choice because of the facile and definednature of the gene locus transfer. The use of Agrobacterium-mediatedplant integrating vectors to introduce DNA into plant cells iswell-known in the art (Fraley et al., Nat. Biotechnol. 3:629-635, 1985;U.S. Pat. No. 5,563,055).

Transformation of plant protoplasts also can be achieved using methodsbased on calcium phosphate precipitation, polyethylene glycol treatment,electroporation, and combinations of these treatments (see, e.g.,Potrykus et al., Mol. Gen. Genet. 199:183-188, 1985; Omirulleh et al.,Plant Mol. Biol. 21(3):415-428, 1993; Fromm et al., Nature 312:791-793,1986; Uchimiya et al., Mol. Gen. Genet. 204:204, 1986; Marcotte et al.,Nature 335:454, 1988). Transformation of plants and expression offoreign genetic elements is exemplified in Choi et al. (Plant Cell Rep.13: 344-348, 1994), and Ellul et al. (Theor. Appl. Genet. 107:462-469,2003).

A number of promoters have utility for plant gene expression for anygene of interest including but not limited to selectable markers,scoreable markers, genes for pest tolerance, disease resistance,nutritional enhancements and any other gene of agronomic interest.Examples of constitutive promoters useful for plant gene expressioninclude, but are not limited to, the cauliflower mosaic virus (CaMV)P-35S promoter, which confers constitutive, high-level expression inmost plant tissues (see, e.g., Odell et al., Nature 313:810, 1985),including in monocots (see, e.g., Dekeyser et al., Plant Cell 2:591,1990; Terada and Shimamoto, Mol. Gen. Genet. 220:389, 1990); a tandemlyduplicated version of the CaMV 35S promoter, the enhanced 35S promoter(P-e35S); the nopaline synthase promoter (An et al., Plant Physiol.88:547, 1988); the octopine synthase promoter (Fromm et al., Plant Cell1:977, 1989); and the figwort mosaic virus (P-FMV) promoter as describedin U.S. Pat. No. 5,378,619 and an enhanced version of the FMV promoter(P-eFMV) where the promoter sequence of P-FMV is duplicated in tandem;the cauliflower mosaic virus 19S promoter; a sugarcane bacilliform viruspromoter; a commelina yellow mottle virus promoter; and other plant DNAvirus promoters known to express in plant cells.

A variety of plant gene promoters that are regulated in response toenvironmental, hormonal, chemical, and/or developmental signals can alsobe used for expression of an operably linked gene in plant cells,including promoters regulated by (1) heat (Callis et al., Plant Physiol.88:965, 1988), (2) light (e.g., pea rbcS-3A promoter, Kuhlemeier et al.,Plant Cell 1:471, 1989; maize rbcS promoter, Schaffner and Sheen, PlantCell 3:997, 1991; or chlorophyll a/b-binding protein promoter, Simpsonet al., EMBO J 4:2723, 1985), (3) hormones, such as abscisic acid(Marcotte et al., Plant Cell 1:969, 1989), (4) wounding (e.g., wunl,Siebertz et al., Plant Cell 1:961, 1989); or (5) chemicals such asmethyl jasmonate, salicylic acid, or Safener. It may also beadvantageous to employ organ-specific promoters (e.g., Roshal et al.,EMBO J 6:1155, 1987; Schernthaner et al., EMBO J 7:1249, 1988; Bustos etal., Plant Cell 1:839, 1989).

Exemplary nucleic acids which may be introduced to plants of thisinvention include, for example, DNA sequences or genes from anotherspecies, or even genes or sequences which originate with or are presentin the same species, but are incorporated into recipient cells bygenetic engineering methods rather than classical reproduction orbreeding techniques. However, the term “exogenous” is also intended torefer to genes that are not normally present in the cell beingtransformed, or perhaps simply not present in the form, structure, etc.,as found in the transforming DNA segment or gene, or genes which arenormally present and that one desires to express in a manner thatdiffers from the natural expression pattern, e.g., to over-express.Thus, the term “exogenous” gene or DNA is intended to refer to any geneor DNA segment that is introduced into a recipient cell, regardless ofwhether a similar gene may already be present in such a cell. The typeof DNA included in the exogenous DNA can include DNA which is alreadypresent in the plant cell, DNA from another plant, DNA from a differentorganism, or a DNA generated externally, such as a DNA sequencecontaining an antisense message of a gene, or a DNA sequence encoding asynthetic or modified version of a gene.

Many hundreds if not thousands of different genes are known and couldpotentially be introduced into a spinach plant according to theinvention. Non-limiting examples of particular genes and correspondingphenotypes one may choose to introduce into a spinach plant include oneor more genes for insect tolerance, such as a Bacillus thuringiensis(B.t.) gene, pest tolerance such as genes for fungal disease control,herbicide tolerance such as genes conferring glyphosate tolerance, andgenes for quality improvements such as yield, nutritional enhancements,environmental or stress tolerances, or any desirable changes in plantphysiology, growth, development, morphology or plant product(s). Forexample, structural genes would include any gene that confers insecttolerance including but not limited to a Bacillus insect control proteingene as described in WO 99/31248, herein incorporated by reference inits entirety, U.S. Pat. No. 5,689,052, herein incorporated by referencein its entirety, U.S. Pat. Nos. 5,500,365 and 5,880,275, hereinincorporated by reference in their entirety. In another embodiment, thestructural gene can confer tolerance to the herbicide glyphosate asconferred by genes including, but not limited to Agrobacterium strainCP4 glyphosate resistant EPSPS gene (aroA:CP4) as described in U.S. Pat.No. 5,633,435, herein incorporated by reference in its entirety, orglyphosate oxidoreductase gene (GOX) as described in U.S. Pat. No.5,463,175, herein incorporated by reference in its entirety.

Alternatively, the DNA coding sequences can affect these phenotypes byencoding a non-translatable RNA molecule that causes the targetedinhibition of expression of an endogenous gene, for example viaantisense- or cosuppression-mediated mechanisms (see, for example, Birdet al., Biotech. Gen. Engin. Rev. 9:207, 1991). The RNA could also be acatalytic RNA molecule (i.e., a ribozyme) engineered to cleave a desiredendogenous mRNA product (see for example, Gibson and Shillito, Mol.Biotech. 7:125, 1997). Thus, any gene which produces a protein or mRNAwhich expresses a phenotype or morphology change of interest is usefulfor the practice of the present invention.

F. DEFINITIONS

In the description and tables herein, a number of terms are used. Inorder to provide a clear and consistent understanding of thespecification and claims, the following definitions are provided:

Allele: Any of one or more alternative forms of a gene locus, all ofwhich alleles relate to one trait or characteristic. In a diploid cellor organism, the two alleles of a given gene occupy corresponding locion a pair of homologous chromosomes.

Backcrossing: A process in which a breeder repeatedly crosses hybridprogeny, for example a first generation hybrid (F₁), back to one of theparents of the hybrid progeny. Backcrossing can be used to introduce oneor more single locus conversions from one genetic background intoanother.

Crossing: The mating of two parent plants.

Cross-pollination: Fertilization by the union of two gametes fromdifferent plants.

Diploid: A cell or organism having two sets of chromosomes.

Emasculate: The removal of plant male sex organs or the inactivation ofthe organs with a cytoplasmic or nuclear genetic factor or a chemicalagent conferring male sterility.

Enzymes: Molecules which can act as catalysts in biological reactions.

F₁ Hybrid: The first generation progeny of the cross of two nonisogenicplants.

Genotype: The genetic constitution of a cell or organism.

Haploid: A cell or organism having one set of the two sets ofchromosomes in a diploid.

Linkage: A phenomenon wherein alleles on the same chromosome tend tosegregate together more often than expected by chance if theirtransmission was independent.

Marker: A readily detectable phenotype, preferably inherited incodominant fashion (both alleles at a locus in a diploid heterozygoteare readily detectable), with no environmental variance component, i.e.,heritability of 1.

Phenotype: The detectable characteristics of a cell or organism, whichcharacteristics are the manifestation of gene expression.

Quantitative Trait Loci (QTL): Quantitative trait loci (QTL) refer togenetic loci that control to some degree numerically representabletraits that are usually continuously distributed.

Resistance: As used herein, the terms “resistance” and “tolerance” areused interchangeably to describe plants that show no symptoms to aspecified biotic pest, pathogen, abiotic influence or environmentalcondition. These terms are also used to describe plants showing somesymptoms but that are still able to produce marketable product with anacceptable yield. Some plants that are referred to as resistant ortolerant are only so in the sense that they may still produce a crop,even though the plants are stunted and the yield is reduced.

Regeneration: The development of a plant from tissue culture.

Royal Horticultural Society (RHS) Colour Chart value: The RHS Colourchart is a standardized reference which allows accurate identificationof any color. A color's designation on the chart describes its hue,brightness and saturation. A color is precisely named by the RHS Colourchart by identifying the group name, sheet number and letter, e.g.,Yellow-Orange Group 19A or Red Group 41B.

Self-pollination: The transfer of pollen from the anther to the stigmaof the same plant.

Single Locus Converted (Conversion) Plant: Plants which are developed bya plant breeding technique called backcrossing, wherein essentially allof the morphological and physiological characteristics of a spinachvariety are recovered in addition to the characteristics of the singlelocus transferred into the variety via the backcrossing technique and/orby genetic transformation.

Substantially Equivalent: A characteristic that, when compared, does notshow a statistically significant difference (e.g., p=0.05) from themean.

Tissue Culture: A composition comprising isolated cells of the same or adifferent type or a collection of such cells organized into parts of aplant.

Transgene: A genetic locus comprising a sequence which has beenintroduced into the genome of a spinach plant by transformation orsite-specific recombination.

G. DEPOSIT INFORMATION

A deposit of spinach hybrid SVVC8155 and inbred parent lineSSBS012-1181F, disclosed above and recited in the claims, has been madewith the American Type Culture Collection (ATCC), 10801 UniversityBlvd., Manassas, Va. 20110-2209. The date of deposit for spinach hybridSVVC8155 and inbred parent line SSBS012-1181F was Mar. 1, 2017. Theaccession numbers for those deposited seeds of spinach hybrid SVVC8155and inbred parent line SSBS012-1181F are ATCC Accession No. PTA-124050and ATCC Accession No. PTA-124049, respectively. Upon issuance of apatent, all restrictions upon the deposits will be removed, and thedeposits are intended to meet all of the requirements of 37 C.F.R. §1.801-1.809. The deposits will be maintained in the depository for aperiod of 30 years, or 5 years after the last request, or for theeffective life of the patent, whichever is longer, and will be replacedif necessary during that period.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity andunderstanding, it will be obvious that certain changes and modificationsmay be practiced within the scope of the invention, as limited only bythe scope of the appended claims.

All references cited herein are hereby expressly incorporated herein byreference.

1. A spinach plant grown from the seed of claim
 2. 2. A spinach seedcomprising at least a first set of the chromosomes of spinach lineSSBS012-1181F, a sample of seed of said line having been deposited underATCC Accession Number PTA-124049. 3-6. (canceled)
 7. The plant of claim1, wherein the hybrid plant is spinach hybrid SVVC8155, a sample of seedof said hybrid SVVC8155 having been deposited under ATCC AccessionNumber PTA-124050.
 8. The seed of claim 2, defined as a seed of spinachhybrid SVVC8155, a sample of seed of said hybrid SVVC8155 having beendeposited under ATCC Accession Number PTA-124050.
 9. (canceled)
 10. Aplant part of the plant of claim
 1. 11. The plant part of claim 10,further defined as a leaf, an ovule, pollen, a fruit, or a cell. 12.(canceled)
 13. A tissue culture of regenerable cells of the plant ofclaim
 1. 14. The tissue culture according to claim 13, comprising cellsor protoplasts from a plant part selected from the group consisting ofembryos, meristems, cotyledons, pollen, leaves, anthers, roots, roottips, pistil, flower, seed and stalks.
 15. A spinach plant regeneratedfrom the tissue culture of claim
 13. 16. A method of vegetativelypropagating the spinach plant of claim 1 comprising the steps of: (a)collecting tissue capable of being propagated from the plant accordingto claim 1; (b) cultivating said tissue to obtain proliferated shoots;and (c) rooting said proliferated shoots to obtain rooted plantlets. 17.The method of claim 16, further comprising growing at least a firstspinach plant from said rooted plantlets.
 18. A method of introducing atrait into a spinach line comprising: (a) utilizing as a recurrentparent a plant of line SSBS012-1181F, by crossing a plant of lineSSBS012-1181F with a second spinach plant that comprises a trait toproduce F1 progeny, a sample of seed of said line having been depositedunder ATCC Accession Number PTA-124049; (b) selecting an F1 progeny thatcomprises the trait; (c) backcrossing the selected F1 progeny with aplant of line SSBS012-1181F to produce backcross progeny; (d) selectingbackcross progeny comprising the trait and the physiological andmorphological characteristics of spinach line SSBS012-1181F; and (e)repeating steps (c) and (d) three or more times to produce selectedfourth or higher backcross progeny that comprises the trait and thephysiological and morphological characteristics of spinach lineSSBS012-1181F.
 19. A spinach plant produced by the method of claim 18.20. A method of producing a spinach plant comprising an added trait, themethod comprising introducing a transgene conferring the trait into aplant of spinach hybrid SVVC8155, or spinach line SSBS012-1181F, asample of seed of said hybrid and line having been deposited under ATCCAccession Number PTA-124050 and ATCC Accession Number PTA-124049,respectively.
 21. (canceled)
 22. A spinach plant comprising at least afirst set of the chromosomes of spinach line SSBS012-1181F, a sample ofseed of said line having been deposited under ATCC Accession NumberPTA-124049, further comprising a transgene.
 23. The plant of claim 22,wherein the transgene confers a trait selected from the group consistingof male sterility, herbicide tolerance, insect resistance, pestresistance, disease resistance, modified fatty acid metabolism,environmental stress tolerance, modified carbohydrate metabolism, andmodified protein metabolism.
 24. A spinach plant comprising at least afirst set of the chromosomes of spinach line SSBS012-1181F, a sample ofseed of said line having been deposited under ATCC Accession NumberPTA-124049, further comprising a single locus conversion.
 25. The plantof claim 24, wherein the single locus conversion confers a traitselected from the group consisting of male sterility, herbicidetolerance, insect resistance, pest resistance, disease resistance,modified fatty acid metabolism, environmental stress tolerance, modifiedcarbohydrate metabolism, and modified protein metabolism.
 26. A methodfor producing a seed of a spinach plant derived from at least one ofspinach hybrid SVVC8155 or spinach line SSBS012-1181F comprising thesteps of: (a) crossing a spinach plant of hybrid SVVC8155, or lineSSBS012-1181F with itself or a second spinach plant; a sample of seed ofsaid hybrid and line having been deposited under ATCC Accession NumberPTA-124050, and ATCC Accession Number PTA-124049, respectively; and (b)allowing seed of a hybrid SVVC8155, or line SSBS012-1181F-derivedspinach plant to form.
 27. The method of claim 26, further comprisingthe steps of: (c) selfing a plant grown from said hybrid SVVC8155, orSSBS012-1181F-derived spinach seed to yield additional hybrid SVVC8155,or line SSBS012-1181F-derived spinach seed; (d) growing said additionalhybrid SVVC8155, or line SSBS012-1181F-derived spinach seed of step (c)to yield additional hybrid SVVC8155, or line SSBS012-1181F-derivedspinach plants; and (e) repeating the crossing and growing steps of (c)and (d) to generate at least a first further hybrid SVVC8155, or lineSSBS012-1181F-derived spinach plant.
 28. (canceled)
 29. The method ofclaim 27, further comprising: (f) crossing the further hybrid SVVC8155,or SSBS012-1181F-derived spinach plant with a second spinach plant toproduce seed of a hybrid progeny plant.
 30. A plant part of the plant ofclaim 7, wherein the plant part comprises a cell of said plant. 31-32.(canceled)
 33. A method of producing a spinach plant part comprising:(a) obtaining the plant according to claim 1, wherein the plant has beencultivated to maturity; and (b) collecting at least a first plant partfrom the plant.
 34. The method of claim 33, wherein the plant partcomprises a leaf.